Cell-associated glycosaminoglycans of human teratocarcinoma-derived cells of line PA 1.
Human teratocarcinoma-derived cells of line PA 1, which are capable of differentiating in vitro [Zeuthen, J. et al. (1980) Int J. Cancer, 25, 19-32], incorporate label from radioactive sulfate and/or glucosamine into several large-sized glycosaminoglycans including hyaluronate, chondroitin sulfate/dermatan sulfate co-polymers, heparan sulfate and keratan-sulfate-like molecules. All these polysaccharide fractions were identified by specific degradation methods. The labeled hyaluronate was degraded into a mixture of unsaturated octa-, hexa- and tetra-saccharides by a treatment with Streptomyces hyaluronidase (EC 4.2.2.1). The chondroitin sulfate/dermatan sulfate co-polymers were cleaved with chondroitin AC lyase (EC 4.2.2.5) into unsaturated disaccharides and a series of unsaturated oligosaccharides; the latter were degraded by a treatment with chondroitin ABC lyase (EC 4.2.2.4) into unsaturated disaccharides. Heparan sulfate was degraded with nitrous acid into free inorganic [35S]sulfate and a series of [35S]sulfate-labeled oligosaccharides and/or glycopeptides. The keratan-sulfate-like molecules were hydrolyzed by a treatment with endo-beta-galactosidase from Escherichia freundii into a series of distinct [35S]sulfate-labeled oligosaccharides; small oligosaccharides were liberated also from [3H]galactose-labeled molecules. The smallest one of the liberated oligosaccharides was tentatively identified as a sulfated disaccharide.
D002460:Cell Line; D002808:Chondroitin Lyases; D002809:Chondroitin Sulfates / Q000032:analysis; D003871:Dermatan Sulfate / Q000032:analysis; D006025:Glycosaminoglycans / Q000032:analysis*; D006026:Glycoside Hydrolases; D006497:Heparan Sulfate / Q000032:analysis; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis; D006821:Hyaluronoglucosaminidase; D007632:Keratan Sulfate / Q000032:analysis; D009608:Nitrous Acid; D013724:Teratoma / Q000032:analysis*; D001616:beta-Galactosidase
D002808:Chondroitin Lyases; D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D006025:Glycosaminoglycans; D006026:Glycoside Hydrolases; D006497:Heparan Sulfate; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase; D007632:Keratan Sulfate; D009608:Nitrous Acid; D001616:beta-Galactosidase; C026093:keratan-sulfate endo-1,4-beta-galactosidase